New Non Nucleosidic Phosphoramidites for the Solid Phase Multi-Labelling of Oligonucleotides: Comb- and Multifork-Like Structures

Vos, Michel J.; Elsen, A Van; Bollen, Alex

doi:10.1080/15257779408013219

Cited by 15 publications

(20 citation statements)

References 26 publications

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“…The PCR-RLBA method was therefore used to analyze 18 cases of neuroborreliosis. The results showed that most patients had been infected with B. garinii (13 out of 18 patients [ Table 4]), in agreement with previous observations showing the preferential association of this genospecies with neurological complications (8,30,27). Moreover, the PCR-RLBA analysis revealed several cases of multiple infection (Table 1).…”

Section: Discussionsupporting

confidence: 79%

“…The condensation of 5Ј phosphate oligonucleotides onto primary amino groups has already been described by De Vos and coworkers (8). Briefly, the 5Ј phosphate oligonucleotides (200 ng/l) were diluted into 10 mM 1-methyl-N-imidazole containing NЈ-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (80 ng/l) and applied to a nylon membrane strip (Biodyne C; Pall Corporation, Brussels, Belgium) with a pump mechanism which delivers controlled amounts of probe to this membrane.…”

Section: Methodsmentioning

confidence: 99%

“…The amplified DNA fragments present sufficient diversity and homology to design genospecific capture probes that allow the typing of each Borrelia genospecies and one detection probe that is able to recognize all tested B. burgdorferi genospecies. The detection probe carries a multifork-like structure on its 5Ј end with eight biotin groups (8). The sequences of both capture and detection probes and their positions within the amplified sequences are shown in Table 1.…”

Section: Methodsmentioning

confidence: 99%

“…RLBA is based on the hybridization of a target DNA, previously amplified by PCR, to a capture probe covalently linked onto the surface of a nylon membrane strip. The hybridized DNA is then recognized by a multiforklike oligonucleotide probe carrying 8 biotin groups that usually increases five times the sensitivity of the assay (8). the PCR products, the nylon strip was incubated for 90 min at 37°C under slight agitation.…”

Section: Methodsmentioning

confidence: 99%

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PCR-Reverse Line Blot Typing Method Underscores the Genomic Heterogeneity of Borrelia valaisiana Species and Suggests Its Potential Involvement in Lyme Disease

Godfroid

Humair

et al. 2003

J Clin Microbiol

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Detection of the Borrelia burgdorferi sensu lato complex in biological samples is currently done by conventional immunological and molecular biological methods. To improve on the accuracy of these methods and to simplify the procedure for testing large numbers of samples, a solid-phase sandwich hybridization system readily applicable to the detection of PCR products has been designed. This colorimetric detection system relies on the use of polybiotinylated detection probes and of specific capture oligonucleotides covalently linked at allocated positions on nylon membrane strips. From a phylogenetic analysis on a great number of ospA gene sequences, we have designed and synthesized a set of PCR primers specific to the five Borrelia burgdorferi sensu lato genospecies present in Europe and a subset of probes (capture and detection probes) specific to these five genospecies (B. burgdorferi sensu stricto, B. garinii, B. afzelii, B. valaisiana, and B. lusitaniae). This combined PCR hybridization system was evaluated with a large number of various B. burgdorferi isolates and clinical specimens. These analyses clearly showed that the system could be used as a typing method to distinguish five genospecies belonging to the B. burgdorferi sensu lato complex. In addition, the study showed that B. valaisiana strains might be more heterologous than suspected up to now and clustered into three genomic groups.

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Section: Discussionsupporting

confidence: 79%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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PCR-Reverse Line Blot Typing Method Underscores the Genomic Heterogeneity of Borrelia valaisiana Species and Suggests Its Potential Involvement in Lyme Disease

Godfroid

Humair

et al. 2003

J Clin Microbiol

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“…To improve the sensitivity and the specificity of the PCR assay, we linked the assay with a colorimetric sandwich capture hybridization assay which has been recently developed for the identification of infectious agents (Lage et al, 1996). The procedure takes advantage of an original DNA chemistry to generate multibiotinylated probes and to bind oligonucleotides onto a plastic support (De Vos, Van Elsen & Bollen, 1994).…”

mentioning

confidence: 99%

Highly specific and sensitive non-radioactive molecular identification of Phytophthora cinnamomi

Coelho¹,

Cravador

Bollen

et al. 1997

Mycological Research

Self Cite

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In response to the need for a faster, more reliable method for identifying Phytophthora cinnamomi in cork oak soils in Portugal, a simple, fast, sensitive molecular identification method is described. It is based on a colorimetric assay which involves an oligonucleotide capture probe covalently immobilised on microtitration wells, a multi-biotinylated oligonucleotide detection probe and the PCR-amplified target DNA. The target DNA is a 349 bp DNA fragment partially covering the 3h-translated and 3h-untranslated regions of the cinnamomin gene. When the specificity of the PCR reaction was evaluated in vitro using isolates of P. cinnamomi and eight other Phytophthora species, including the related P. cambivora, it was specific to P. cinnamomi. When 30 isolates of P. cinnamomi from oak roots in southern Portugal were assayed, 26 gave a strong positive response. The assay has a sensitivity of about 2-5 genome equivalents of P. cinnamomi. The reason for the negative response of four isolates remains unclear.

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Solid‐Phase Supports for Oligonucleotide Synthesis

Guzaev¹

2013

CP Nucleic Acid Chemistry

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This unit attempts to provide a reasonably complete inventory of over 280 solid supports available to oligonucleotide chemists for preparation of natural and 3'-modified oligonucleotides. Emphasis is placed on non-nucleosidic solid supports. The relationship between the structural features of linkers and their behavior in oligonucleotide synthesis and deprotection is discussed wherever the relevant observations are available.

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New Non Nucleosidic Phosphoramidites for the Solid Phase Multi-Labelling of Oligonucleotides: Comb- and Multifork-Like Structures

Cited by 15 publications

References 26 publications

PCR-Reverse Line Blot Typing Method Underscores the Genomic Heterogeneity of Borrelia valaisiana Species and Suggests Its Potential Involvement in Lyme Disease

PCR-Reverse Line Blot Typing Method Underscores the Genomic Heterogeneity of Borrelia valaisiana Species and Suggests Its Potential Involvement in Lyme Disease

Highly specific and sensitive non-radioactive molecular identification of Phytophthora cinnamomi

Solid‐Phase Supports for Oligonucleotide Synthesis

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